CN107698575A - The bifunctional molecule of novel B ET protein degradations ligand-mediated cereblon and its preparation and application - Google Patents
The bifunctional molecule of novel B ET protein degradations ligand-mediated cereblon and its preparation and application Download PDFInfo
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Abstract
The present invention relates to new difunctional small molecule and its pharmaceutically acceptable salt, the hydrate or preparation method of prodrug and these compounds and its Pharmaceutical composition in treatment tumour, inflammation, the application in disease such as is immunized.Difunctional small molecule of the present invention is a kind of protein degradation targeting association (PROTACs), and they being capable of selective induction BET protein degradations.Difunctional small molecule is obtained by the present invention in that being connected BET protein micromolecular inhibitors with cereblon protein ligands in E3 SCF-complexes with linking arm.
Description
Technical field
The present invention relates to the preparation method of new difunctional small molecule and its pharmaceutically acceptable salt, hydrate or prodrug
And these compounds and its Pharmaceutical composition be in treatment tumour, inflammation, the application in disease such as it is immunized.It is of the present invention double
Function small molecule is a kind of protein degradation targeting association (PROTACs), and they being capable of selective induction BET protein degradations.This
Invention is by using linking arm by cereblon protein ligands in BET protein micromolecular inhibitors and E3 SCF-complexes
Connection obtains difunctional small molecule.
Background technology
The ubiquitination of target protein generally requires the collective effect of three enzymes, is E1 activating enzymes first to activate ubiquitin molecule,
Then E2 desmoenzymes carry out the ubiquitin molecule of " clasping " activation, finally will be general by the E3 ubiquitin ligases and E2 of binding specificity substrate
Fibroin is covalently transferred on one or more lysine residues of target protein.This mechanism of degradation to specific protein high special
What system was mainly determined by E3 ubiquitin ligases, therefore E3 ligases are conclusive central angle in the degradation process of target protein
Color.E3 ubiquitin ligases containing Ring domains are a huge E3 ubiquitin ligase monoids, are responsible for probably about 20%
Proteins ubiquitinization is degraded, and the degraded event that the Cullin-Ring Ligase (CRL) being made up of Cullin albumen etc. are regulated and controled,
Occupy sizable ratio wherein.The E3 complexs that DDB1, CUL4 and ROC1 are formed are that an attested ubiquitination connects
Enzyme is connect, DNA reparation, duplication and transcription can be adjusted.Research finds that other many factors also assist in E3 proteasome complex
Formation, these factors are the CUL4 correlation factors of DDB1 mono- by Uniform Name, i.e. DCAF.Although CRBN is presently believed to be participation
The correlation factor that E3 compounds are formed, but its major function in the complex is unclear.Cereblon ubiquitination target eggs
Cause FGF8 (FGF8) and Fibroblast growth factor-10 (FGF10) to increase in vain, illustrate ubiquitination enzyme
Complex grows particularly significant for fetal mouse limb.
Lenalidomide is used to treat immunological diseases and tumour, including Huppert's disease.Research shows that lenalidomide can be with
With reference to cereblon, so as to trigger transcription factor Ikaros (IKZF1) and Aiolos (IKZF3) ubiquitination and degraded, these
Growth of the transcription factor for Huppert's disease is of crucial importance.In the treatment of current Huppert's disease, height expression
The effect of cereblon is with lenalidomide or similar medicine is closely related.
BET protein families are considered as the potential target spot of numerous diseases especially cancer, obtain many research institutes and medicine enterprise
Concern.BET family proteins include four member proteins BRD2, BRD3, BRD4 and BRDT, and each albumen includes two independences
Bromodomain (BD1, BD2.Wherein, BRD2, BRD3 and BRD4 are widely distributed, can almost be sent out in all cells of human body
It is existing, and BRDT is only distributed in testis.Bromodomain domains are one highly conserved by about 110 Amino acid profiles
Protein function domain, albumen are made up of four α spirals (Z, A, B, C) and two Loop (ZA, BC), can form hydrophobic region
Domain, identify and combine the lysine residue of acetylation.
BRD4 plays a significant role in regulatory gene transcription, many studies have shown that BRD4 is raised and arrived target gene
Super enhancer region, promote the transcription of the oncogene such as c-MYC, Bcl-X1 and BCL-6.Because BRD4 is in regulation oncogene turn
Key effect in record, potential anti-tumor target is become, combined using BET inhibitor interference BRD4 with target gene, under
The oncogene such as c-MYC are adjusted, reach antitumor purpose.
In recent years, BET protein inhibitors are by domestic and international researcher's extensive concern, and lot of documents and patent are to BET eggs
White inhibitor is reported.Up to now, domestic and international pharmacy corporation, colleges and universities and research institution apply for that more than 200 pieces are related to BET altogether
The patent of protein inhibitor.Although successfully being listed still without BET inhibitor, share 16 drug candidates and be in the I/II phases and face
Bed conceptual phase, principal indication is malignant tumour.Although BET inhibitor is shown in numerous experiment in vitro and animal model
Good active anticancer and tolerance, but clinical research still finds there are problems that.
The difunctional small molecule designed in the present invention can carry out ubiquitination mark to BRD4 albumen, and inducible protein is degraded,
Antitumous effect is better than BET inhibitor.Suppress BRD4 albumen to generally require to maintain medicine into higher concentration for a long time, it is possible to
Cause serious side effect；And inducible protein degraded only needs a small amount of medicine can, this process is similar to catalytic reaction,
And the medicine of equimolar amounts is not needed, so medicine dosage can be reduced using difunctional small molecule, mitigate toxic side effect.
The content of the invention
It is an object of the invention to provide some new difunctional small molecules and its pharmaceutically acceptable salt, hydrate or
Prodrug.These compounds have the function of induction BET protein degradations, available for preparing new type antineoplastic medicine.The tumour can be
But it is not limited to the white blood of Huppert's disease, stomach cancer, lung cancer, breast cancer, the cancer of the esophagus, colon cancer, medulloblastoma, acute myelogenous
Disease, chronic leukemia, prostate cancer, hepatoma, renal cell carcinoma, cervical carcinoma, cutaneum carcinoma, oophoroma, colon cancer, neuroglia
Knurl, thyroid cancer or cancer of pancreas.
The present invention also aims to provide a kind of preparation method for synthesizing new difunctional small molecule.
Another object of the present invention is to provide a kind of pharmaceutical preparation containing new difunctional small molecule.
Detailed description content is as follows：
To achieve these goals, the invention provides the difunctional small molecule shown in below formula or its can pharmaceutically connect
Salt, hydrate or the prodrug received：
A-L-B
Wherein：
A is the smaller ligand of cereblon albumen in E3 SCF-complexes, comprising amides compound, adjacent benzene
Diimide compounds, Thalidomide or derivatives thereof, lenalidomide or derivatives thereof, pomalidomide or derivatives thereof, portion
Dispensing body general structure is as follows：
Wherein：
W is selected from CH2, C=O, SO2, NH, N- alkyl；
X is selected from O, S；
Y is selected from NH, N- alkyl, N- aryl, N- heterocycles, N- cycloalkyl, O, S；
Z be selected from-alkyl ,-cycloalkyl ,-Cl ,-F；
G, G ' is selected from-H ,-alkyl ,-OH ,-CH2- heterocycle；
Q1、Q2、Q3、Q4Selected from C, N, NO.
L is linking arm, comprising non-linear chain, aliphatic chain, fragrant chain, Heteroaryl ring structures chain, passes through covalent bond and A and B
It is connected, part is connected shown in arm configuration below formula：
Wherein：
N is selected from the integer between 1-10.
B is BET protein inhibitors or derivatives thereof.
Present invention is preferably related to the difunctional small molecule shown in below formula (Ia), (IIa) and (IIIa) or its pharmaceutically
Acceptable salt, hydrate or prodrug：
Wherein：
R1Selected from-H ,-D ,-CH3、-CH2F、CHF2、-CF3、-CH2D、-CHD2、-CD3、-CH2CH3、-CH2CH2CH3；
R2、R3It is identical or different, separately selected from-H ,-D ,-F ,-Cl ,-Br ,-I ,-NO2、-CN、-NH2、-OH、-
CH3、-CH2F、CHF2、-CF3、-CH2D、-CHD2、-CD3、-CH2CH3；
N is selected from the integer between 1-10.
The present invention, which is more preferably related to difunctional small molecule shown in formula (Ia), (IIa) and (IHa) or its, can pharmaceutically connect
Salt, hydrate or the prodrug received：
Wherein：
R1Selected from-H ,-CH3、-CH2CH3；
R2、R3It is identical or different, separately selected from-H ,-F ,-Cl ,-Br ,-CH3、-CH2F、CHF2、-CF3、-
CH2D、-CHD2、-CD3；
N is selected from the integer between 1-6.
Preferred compounds of the invention includes, but are not limited to：
Ia class compounds：
2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydro quinoline azoles
Quinoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl)-N- (2- (2,6- dioxopiperidine -3- bases) -1-
Oxoisoindolines -4- bases) acetamide；
2- (2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydro
Quinazoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl) ethyoxyl)-N- (2- (2,6- dioxy piperazines
Pyridine -3- bases) -1- oxoisoindolines -4- bases) acetamide；
2- (2- (2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-dihydro quinoline
Oxazoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl) ethyoxyl) ethyoxyl)-N- (2- (2,6- dioxies
For piperidines -3- bases) -1- oxoisoindolines -4- bases) acetamide；
2- (2- (2- (2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxo -4- phenyl-two
Hydrogen quinazoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl) ethyoxyl) ethyoxyl) ethyoxyl)-N-
(2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) acetamide；
IIa class compounds：
3- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) propionamide；
4- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) butyramide；
5- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) pentanamide；
6- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) caproamide；
7- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) heptamide；
8- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) caprylamide；
IIIa class compounds
3- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) propionamide；
5- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) pentanamide；
6- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) caproamide；
7- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) heptamide；
The structure of part of compounds is：
Ia class compounds：
IIa class compounds：
IIIa class compounds
Compound shown in formula (Ia), (IIa) and (IIIa) can contain asymmetric or chiral centre, therefore can be with
Different stereoisomeric forms in any ratio are present.All stereoisomeric forms in any ratio of the compounds of this invention, including but not limited to diastereoisomer,
Enantiomter and atropisomer and their mixture (such as racemate), are included within the scope of the present invention.
Compound shown in formula (Ia), (IIa) and (IIIa) can also exist with different tautomeric forms, it is all this
A little forms are included in the scope of the invention.Belong to " dynamic isomer " or " tautomeric form " to refer to build mutually via low energy
The constitutional isomer of the different-energy of conversion.
According to the present invention, pharmaceutically acceptable salt includes the addition salts formed with following acid：Hydrochloric acid, hydrobromic acid, sulfuric acid,
Phosphoric acid, methanesulfonic acid, ethyl sulfonic acid, p-methyl benzenesulfonic acid, benzene sulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid,
Citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid etc..Hydrochloric acid, hydrobromic acid, sulfuric acid, citric acid, tartaric acid, phosphoric acid, lactic acid,
Pyruvic acid, acetic acid, trifluoroacetic acid, maleic acid, benzene sulfonic acid, butanedioic acid and similar known acceptable acid are into salt.
In addition, present invention additionally comprises the prodrug of derivative of the present invention.The prodrug of derivative of the present invention be formula (Ia),
(IIa) and (IIIa) derivative, their own may have weaker activity even without activity, but upon administration,
(such as by metabolism, solvolysis or other mode) is converted to corresponding biologically active form under physiological condition.
The present invention can contain the difunctional small molecule of above formula (Ia), (IIa) and (IIIa) and its pharmaceutically acceptable
Salt, hydrate are prepared by mixing into composition, and be prepared into and clinically may be used as active ingredient with pharmaceutically acceptable excipient
The formulation of receiving, above-mentioned excipient refer to diluent, adjuvant or the carrier available for pharmaceutical field.Above-mentioned formulation refers to clinic
Upper conventional injection, tablet, capsule etc..
Compound of the present invention or its pharmaceutically acceptable salt, hydrate, prodrug can be used as unique antitumor
Medicine is used alone, or can be used in combination with the antineoplastic listed, for Prevention tumour etc..
Examples provided hereinafter and preparation example further elucidate and illustrated the compounds of this invention and its preparation side
Method.It should be appreciated that the scope that the scope of following examples and preparation example is not limit the invention in any way.
The compounds process for production thereof of formula (Ia), method are as follows：
Step a：Compound I reacts to obtain compound II with paratoluensulfonyl chloride；
Step b：Compound II reacts to obtain compound III with Sodium azide；
Step c：Compound III reacts to obtain compound IV with bromo-acetic acid tert-butyl；
Step d：Compound IV reacts to obtain compound V with trifluoroacetic acid；
Step e：Compound V reacts to obtain compound VI with thionyl chloride；
Step f：Compound VI and lenalidomide or derivatives thereof reaction obtain compound VII；
Step g：Compound VIII and VII reacts to obtain the compound of formula (Ia) expression；
The compounds process for production thereof of formula (IIa), method are as follows：
Step h：Compound IX and lenalidomide or derivatives thereof reaction obtain compound X；
Step i：Compound X reacts to obtain compound XI with trifluoroacetic acid；
Step j：Compound XII and XI reacts to obtain the compound of formula (IIa) expression；
The compounds process for production thereof of formula (IIIa), method are as follows：
Step k：Compound XIH and XVI reacts to obtain compound XV；
Step l：Compound XV and lenalidomide or derivatives thereof reaction obtain the compound of formula (IIIa) expression；
Wherein, R1、R2、R3It is as defined above with n.
Embodiment
It is not required to be further described, it is believed that those skilled in the art, can be at utmost by description above
Using the present invention.Therefore, examples provided below is only that the present invention is furture elucidated, is not meant to any
Mode limits the scope of the invention.
Raw material can obtain from commercial channels, or be prepared by methods known in the art, or according to side described herein
It is prepared by method.
The structure of compound by nuclear magnetic resonance (1H-NMR) and/or mass spectrum (MS) determines.NMR measure is to use ACF-
300BRUK type NMRs, measure solvent is deuterochloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-D6), TMS are interior
Mark.MS measure HP1100 type mass spectrographs.Column chromatography uses 200-300 mesh silica gel (Haiyang Chemical Plant, Qingdao's production).
Ia class compound synthesis
Embodiment 1：Preparation 2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxo -4- phenyl -
Isosorbide-5-Nitrae-dihydroquinazoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl)-N- (2- (2,6- dioxy piperazines
Pyridine -3- bases) -1- oxoisoindolines -4- bases) acetamide；(I-1), its structural formula is as follows：
Step 1) 2- hydroxyethyl -4- toluene sulfonic acides esters (1a)
Ethylene glycol (4.00g, 64.05mmol) is dissolved in 5mL pyridines, be added portionwise paratoluensulfonyl chloride (6.14g,
32.22mmol), after being stirred at room temperature 4 hours, 6mol/L hydrochloric acid (40mL) is added, is extracted with ethyl acetate, saturated common salt washing,
Collected organic layer, anhydrous sodium sulfate drying, decompression boil off organic solvent, and residue is purified by silica gel column chromatography, use
Petrol ether/ethyl acetate (V/V=20/1-10/1) affords colourless liquid, weight 2.34g, yield 33.58%.
1H NMR (300MHz, CDCl3) δ 7.81 (d, J=8.3Hz, 2H), 7.36 (d, J=8.0Hz, 2H), 4.16-4.12
(m, 2H), 3.85-3.78 (m, 2H), 2.45 (s, 3H), 2.01 (d, J=19.0Hz, 1H).
Step 2) 2- hydroxyethyls nitrine (1b)
1a (2g, 9.25mmol) and sodium azide (1.2g, 18.5mmol) are dissolved in 5mL acetone and 5mL water mixed liquids,
It is heated to reflux adding 20mL water in 16 hours, is extracted with ethyl acetate, saturated aqueous sodium carbonate is washed, and saturated common salt washing, is collected
Organic layer, anhydrous sodium sulfate drying, decompression boil off organic solvent, obtain colourless liquid, weight 0.73g, yield 90.64%.
1H NMR (300MHz, CDCl3) δ 3.82-3.74 (m, 2H), 3.49-3.39 (m, 2H), 2.01 (s, 1H).
Step 3) the 2- nitrine ethoxyacetic acid tert-butyl esters (1c)
1b (0.65g, 7.46mmol) and bromo-acetic acid tert-butyl (1.6g, 8.21mmol) are dissolved in 15mL tetrahydrofurans,
Stirred 10 minutes in ice-water bath, sodium hydride (0.53g, 22.39mmol) is added portionwise, it is small to continue the reaction 0.5 in ice-water bath
Shi Hou, react at room temperature 3 hours, add 30mL water, be extracted with ethyl acetate, saturated common salt washing, collected organic layer, anhydrous slufuric acid
Sodium is dried, and decompression boils off organic solvent, and residue is purified by silica gel column chromatography, uses petrol ether/ethyl acetate (V/V
=30/1-20/1) afford colourless liquid, weight 0.43g, yield 28.63%.
1H NMR (300MHz, CDCl3) δ 4.06-4.00 (m, 2H), 3.76-3.70 (m, 2H), 3.48-3.41 (m, 2H),
1.49 (d, J=5.8Hz, 9H).
Step 4) 2- nitrine ethoxyacetic acid (1d)
1c (0.43g, 2.14mmol) is dissolved in 4mL dichloromethane, adds 1mL trifluoroacetic acids, is reacted at room temperature 3 hours
Afterwards, decompression boils off organic solvent, obtains brown liquid, weight 0.29g.
Step 5) 2- nitrine Ethoxyacetyl chloride (1e)
1d (0.29g, 2.03mmol) is dissolved in 4mL dichloromethane, 1mL thionyl chlorides is added, adds 2 drop DMF and urge
Change, after being heated to reflux 4 hours, decompression boils off organic solvent, obtains brown liquid, weight 0.33g.
Step 6) 2- (2- nitrine ethyoxyl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4-
Base) acetamide (1f)
Lenalidomide (0.26g, 0.95mmol) is dissolved in 3mL 1-METHYLPYRROLIDONEs, addition 1e (0.32g,
1.9mmol), after reacting at room temperature 4 hours, 20mL water is added, there is solid precipitation, filtered, dry crude product, pass through silica gel column chromatography
Chromatographic purifying, yellow solid, weight 0.21g, yield are afforded using methylene chloride/methanol (V/V=150/1-100/1)
55.13%.
Step 7) 2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-two
Hydrogen quinazoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl)-N- (2- (2,6- dioxopiperidine -3-
Base) -1- oxoisoindolines -4- bases) acetamide (I-1)
By 6- (3,5- dimethyl isoxazole -4- bases) -1- methyl 4-phenyls -3- (propyl- 2- alkynes -1- bases) -3,4- dihydro quinolines
Oxazoline -2 (1H) -one (0.06g, 0.161mmol), 1f (0.064g, 0.161mmol) and Salzburg vitriol (0.008g,
0.032mmol) it is dissolved in 2mL dichloromethane, in 2mL methanol and 2mL water mixed liquids, is stirred at room temperature 10 minutes, adds ascorbic acid
Sodium (0.013g, 0.065mmol), after reacting at room temperature 4 hours, 20mL water is added, is extracted with dichloromethane, saturated common salt washing,
Collected organic layer, anhydrous sodium sulfate drying, decompression boil off organic solvent, and residue is purified by silica gel column chromatography, use
Methylene chloride/methanol (V/V=100/1-30/1) affords yellow solid, weight 0.057g, yield 45.72%.
MS (ESI, m/z)：757.30[M-H]-
1H NMR (300MHz, CDCl3) δ 8.81 (d, J=34.0Hz, 1H), 8.40 (s, 1H), 8.17 (s, 1H), 7.69
(d, J=7.5Hz, 1H), 7.50 (d, J=7.1Hz, 1H), 7.31 (s, 2H), 7.24 (d, J=4.9Hz, 3H), 7.10 (d, J=
8.0Hz, 1H), 6.93 (d, J=7.9Hz, 2H), 5.70 (s, 1H), 5.30-5.16 (m, 1H), 5.02 (s, 1H), 4.41 (d, J
=70.1Hz, 5H), 3.98 (s, 2H), 3.43 (s, 3H), 2.82 (s, 3H), 2.39-2.02 (m, 7H), 1.78 (s, 3H).
Embodiment 2：Prepare 2- (2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxo -4- benzene
Base-Isosorbide-5-Nitrae-dihydroquinazoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl) ethyoxyl)-N- (2- (2,
6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) acetamide (I-2), its structural formula is as follows：
Synthesis step is the same as embodiment 1
MS (ESI, m/z)：800.00[M-H]-
1H NMR (300MHz, CDCl3) δ 9.18 (d, J=78.9Hz, 1H), 8.63 (d, J=14.2Hz, 1H), 7.81
(dd, J=13.0,7.8Hz, 2H), 7.69 (d, J=7.5Hz, 1H), 7.45 (dd, J=13.2,7.5Hz, 1H), 7.30 (dd, J
=13.1,5.4Hz, 5H), 7.06 (d, J=8.5Hz, 1H), 6.88 (dd, J=8.2,4.1Hz, 2H), 5.75 (d, J=
10.6Hz, 1H), 5.22 (d, J=9.3Hz, 1H), 5.13-4.99 (m, 1H), 4.51-4.30 (m, 4H), 4.07 (dt, J=
15.2,8.7Hz, 3H), 3.87 (s, 2H), 3.76-3.56 (m, 4H), 3.38 (d, J=2.0Hz, 3H), 2.82 (dd, J=
11.9,4.6Hz, 2H), 2.30 (t, J=17.4Hz, 4H), 2.15 (t, J=12.0Hz, 4H), 1.96 (s, 1H).
Embodiment 3：Preparation 2- (2- (2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -
4- phenyl-dihydroquinazoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl) ethyoxyl) ethyoxyl) -
N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) acetamide (I-3), its structural formula are as follows：
Synthesis step is the same as embodiment 1
MS (ESI, m/z)：844.05[M-H]-
1H NMR (300MHz, CDCl3) δ 9.17 (d, J=49.8Hz, 2H), 7.75 (d, J=7.0Hz, 1H), 7.63 (s,
1H), 7.39 (dd, J=55.8,14.0Hz, 6H), 7.08 (d, J=7.9Hz, 1H), 6.90 (s, 2H), 5.82 (s, 1H), 5.23
(s, 1H), 4.51-4.12 (m, 6H), 3.60 (dd, J=62.3,41.9Hz, 14H), 2.87 (s, 2H), 2.25 (t, J=
34.5Hz, 9H), 1.81 (s, 2H).
Embodiment 4：Prepare 2- (2- (2- (2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxygen
Generation -4- phenyl-dihydroquinazoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl) ethyoxyl) ethoxy
Base) ethyoxyl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) acetamide (I-4), its structure
Formula is as follows：
Synthesis step is the same as embodiment 1
MS (ESI, m/z)：888.05[M-H]-
1H NMR (300MHz, CDCl3) δ 9.36 (d, J=24.8Hz, 1H), 9.17 (s, 1H), 7.88-7.62 (m, 3H),
7.48 (d, J=7.6Hz, 1H), 7.34 (s, 4H), 7.07 (d, J=8.1Hz, 1H), 6.89 (d, J=12.3Hz, 2H), 5.79
(s, 1H), 5.17 (d, J=14.4Hz, 2H), 4.45 (s, 4H), 4.13 (s, 2H), 4.05 (d, J=16.7Hz, 1H), 3.95-
3.02 (m, 18H), 2.87 (s, 2H), 2.57-2.02 (m, 9H), 1.97 (s, 1H)
IIa class compound synthesis
Embodiment 5：Prepare 3- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitraes -
Dihydroquinazoline -3 (2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases)
Propionamide (II-1), its structural formula are as follows：
Step 1) the tert-butyl group (3- ((2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) amino) -3-
Propionyl) carbamate (2a)
Lenalidomide (0.2g, 0.77mmol), 3- ((tertbutyloxycarbonyl) amino) propionic acid (0.16g, 0.85mmol) is molten
In 2mL acetonitriles, pyridine (0.18g, 2.31mmol) is added, the POCl3 of 2ml dilution in acetonitrile is added under condition of ice bath
(0.14g, 0.92mmol), it is added dropwise and removes ice bath, after reacting at room temperature 4 hours, adds 20mL water, be extracted with ethyl acetate,
Saturated common salt is washed, collected organic layer, anhydrous sodium sulfate drying, and decompression boils off organic solvent, and residue passes through silica gel column chromatography
Chromatographic purifying, faint yellow solid, weight 0.16g, yield are afforded using methylene chloride/methanol (V/V=100/1-40/1)
48.18%.
Step 2) 3- amino-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) propionamide
(2b)
2a (0.16g, 0.37mmol) is dissolved in 4mL dichloromethane, adds 1mL trifluoroacetic acids, is reacted at room temperature 3 hours
Afterwards, decompression boils off organic solvent, obtains yellow solid, weight 0.13g.
Step 3) 3- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydro quinolines
Oxazoline -3 (2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) propionamide
(II-1)
By 2b (0.12g, 0.36mmol) and 2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxo -4- benzene
Base-Isosorbide-5-Nitrae-dihydroquinazoline 3 (2H)-yl) acetic acid (0.12g, 0.30mmol) is dissolved in 2mlDMF, add N- ethyl diisopropylamines
(0.12g, 0.91mmol) adds 2- (7- aoxidizes BTA)-N, N, N ', N '-tetramethylurea hexafluoro phosphorus after reacting 10 minutes
Acid esters (0.14g, 0.36mmol), after reacting at room temperature 4 hours, 20mL water is added, is extracted with dichloromethane, saturated common salt washing,
Collected organic layer, anhydrous sodium sulfate drying, decompression boil off organic solvent, and residue is purified by silica gel column chromatography, use
Methylene chloride/methanol (V/V=100/1-30/1) affords yellow solid, weight 0.152g, yield 71.35%.
MS (ESI, m/z)：702.00[M-H]-
1H NMR (300MHz, DMSO) δ 11.02 (s, 1H), 9.85 (s, 1H), 8.11 (s, 1H), 7.80 (d, J=
7.0Hz, 1H), 7.53-7.43 (m, 2H), 7.36-7.23 (m, 6H), 7.18 (d, J=11.0Hz, 1H), 7.07 (d, J=
8.5Hz, 1H), 5.65 (d, J=3.4Hz, 1H), 5.14 (d, J=8.4Hz, 1H), 4.38 (d, J=8.4Hz, 2H), 3.33 (s,
8H), 3.17 (s, 2H), 2.54 (s, 3H), 2.31 (d, J=2.8Hz, 3H), 2.14 (d, J=1.9Hz, 3H).
Embodiment 6：Prepare 4- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitraes -
Dihydroquinazoline -3 (2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases)
Butyramide (II-2)；Its structural formula is as follows：
Synthesis step is the same as embodiment 5
MS (ESI, m/z)：716.05[M-H]-
1H NMR (300MHz, DMSO) δ 11.03 (s, 1H), 9.82 (s, 1H), 8.03 (s, 1H), 7.81 (d, J=
6.8Hz, 1H), 7.50 (d, J=7.4Hz, 2H), 7.36 (s, 4H), 7.29 (d, J=8.1Hz, 3H), 7.10 (d, J=8.5Hz,
1H), 5.69 (s, 1H), 5.15 (dd, J=13.3,4.8Hz, 1H), 4.38 (d, J=5.6Hz, 2H), 3.35 (s, 7H), 3.16
(s, 2H), 2.58 (d, J=29.7Hz, 3H), 2.44-2.29 (m, 6H), 2.17 (d, J=1.4Hz, 3H).
Embodiment 7：Prepare 5- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitraes -
Dihydroquinazoline -3 (2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases)
Pentanamide (II-3)；Its structural formula is as follows：
Synthesis step is the same as embodiment 5
MS (ESI, m/z)：730.05[M-H]-
1H NMR (300MHz, DMSO) δ 11.01 (s, 1H), 9.77 (s, 1H), 7.96 (s, 1H), 7.80 (d, J=
7.5Hz, 1H), 7.47 (d, J=8.2Hz, 2H), 7.34 (d, J=4.3Hz, 4H), 7.26 (d, J=7.2Hz, 3H), 7.07 (d,
J=9.0Hz, 1H), 5.66 (s, 1H), 5.18-5.08 (m, 1H), 4.40 (dd, J=26.9,9.8Hz, 4H), 3.30 (s, 3H),
2.36 (s, 2H), 2.30 (s, 3H), 2.14 (s, 3H), 1.98 (s, 2H), 1.51 (d, J=47.6Hz, 6H), 1.23 (s, 2H).
Embodiment 8：Prepare 6- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitraes -
Dihydroquinazoline -3 (2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases)
Caproamide (II-4)；Its structural formula is as follows：
Synthesis step is the same as embodiment 5
MS (ESI, m/z)：744.15[M-H]-
1H NMR (300MHz, CDCl3) δ 8.91 (d, J=17.4Hz, 1H), 8.42 (s, 1H), 7.76-7.46 (m, 3H),
7.29 (d, J=32.1Hz, 5H), 7.08 (d, J=8.8Hz, 1H), 6.90 (d, J=8.3Hz, 1H), 6.80 (s, 1H), 6.48
(s, 1H), 5.58 (s, 1H), 5.01 (s, 1H), 4.30 (s, 2H), 3.03 (s, 2H), 2.67 (s, 3H), 2.27 (s, 2H), 2.08
(s, 3H), 1.65 (s, 8H), 1.26 (d, J=39.9Hz, 7H).
Embodiment 9：Prepare 7- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitraes -
Dihydroquinazoline -3 (2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases)
Heptamide (II-5)；Its structural formula is as follows：
MS (ESI, m/z)：758.10[M-H]-
1H NMR (300MHz, DMSO) δ 11.00 (s, 1H), 9.75 (s, 1H), 7.82 (dd, J=15.5,10.5Hz,
2H), 7.49 (t, J=6.7Hz, 2H), 7.35 (d, J=4.0Hz, 4H), 7.27 (d, J=9.1Hz, 3H), 7.08 (d, J=
8.2Hz, 1H), 5.66 (s, 1H), 5.14 (dd, J=13.1,5.1Hz, 1H), 4.36 (d, J=4.1Hz, 2H), 3.31 (s,
5H), 3.05 (d, J=6.3Hz, 2H), 2.36 (d, J=7.3Hz, 2H), 2.32 (s, 3H), 2.15 (s, 3H), 2.01 (s, 2H),
1.60 (s, 2H), 1.33 (dd, J=33.6,12.8Hz, 8H).
Embodiment 10：Preparation 8- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxo -4- phenyl -1,
4- dihydroquinazolines -3 (2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4-
Base) caprylamide (II-6)；Its structural formula is as follows：
MS (ESI, m/z)：772.05[M-H]-
1H NMR (300MHz, DMSO) δ 11.00 (s, 1H), 9.74 (s, 1H), 7.87 (s, 1H), 7.82 (d, J=
6.8Hz, 1H), 7.49 (d, J=7.1Hz, 2H), 7.35 (d, J=3.9Hz, 4H), 7.27 (d, J=10.7Hz, 3H), 7.08
(d, J=8.3Hz, 1H), 5.66 (s, 1H), 5.14 (dd, J=13.1,4.9Hz, 1H), 4.34 (t, J=10.8Hz, 2H),
3.28 (d, J=20.6Hz, 5H), 3.04 (d, J=5.7Hz, 2H), 2.40-2.34 (m, 2H), 2.32 (s, 3H), 2.16 (s,
3H), 2.03 (d, J=9.4Hz, 2H), 1.60 (s, 2H), 1.37 (d, J=6.2Hz, 2H), 1.26 (d, J=12.7Hz, 8H)
Embodiment 11：Prepare 3- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-two
Hydrogen quinazoline -3 (2H)-yl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) propionamide (III-
1)；Its structural formula is as follows：
Step 1) 3- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydro quinoline azoles
Quinoline -3 (2H)-yl) propionic acid (3a)
By 6- (3,5- dimethyl isoxazole -4- bases) -1- methyl 4-phenyls -3,4- dihydroquinazoline -2 (1H) -one
(0.2g, 0.60mmol), 3- bromo-propionic acids (0.14g, 0.90mmol), is dissolved in 2mL DMF, and sodium hydrogen is added under condition of ice bath
(0.029g, 1.2mmol), half an hour recession removes ice bath, after reacting at room temperature 4 hours, adds 20mL water, is extracted with ethyl acetate,
Saturated common salt is washed, collected organic layer, anhydrous sodium sulfate drying, and decompression boils off organic solvent, and residue passes through silica gel column chromatography
Chromatographic purifying, faint yellow solid, weight 0.16g, yield 65.78% are afforded using petrol ether/ethyl acetate (V/V=4/1).
Step 2) 3- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydro quinoline azoles
Quinoline -3 (2H)-yl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) propionamide (III-1)
3a (0.16g, 0.40mmol) and lenalidomide (0.11g, 0.43mmol) are dissolved in 4mL acetonitriles, add pyridine
(0.09g, 1.18mmol), it is slowly added dropwise under condition of ice bath and is dripped with the POCl3 (0.07g, 0.47mmol) of 2ml dilution in acetonitrile
After room temperature condition reaction being transferred to after adding 3 hours, add a small amount of water decompression to boil off acetonitrile, add 20mL water, use ethyl acetate
Extraction, saturated common salt washing, collected organic layer, anhydrous sodium sulfate drying, decompression boil off organic solvent, and residue passes through silicagel column
Chromatographic purification, faint yellow solid is afforded using methylene chloride/methanol (V/V=100/1-20/1), weight 0.08g, is received
Rate 31.35%.
MS (ESI, m/z)：645.25[M-H]-
1H NMR (300MHz, DMSO) δ 10.98 (s, 1H), 9.86 (s, 1H), 7.70 (d, J=7.3Hz, 1H), 7.45
(d, J=6.8Hz, 1H), 7.34 (d, J=4.1Hz, 4H), 7.22 (d, J=10.5Hz, 2H), 7.02 (d, J=8.5Hz, 1H),
6.90 (d, J=7.5Hz, 1H), 6.78 (d, J=7.8Hz, 1H), 5.41 (s, 2H), 3.29 (s, 2H), 2.91 (s, 3H), 2.62
(s, 3H), 2.29 (d, J=4.4Hz, 4H), 2.12 (d, J=4.3Hz, 2H), 1.36-1.27 (m, 1H), 1.22 (s, 3H),
0.84 (s, 1H)
Embodiment 12：Prepare 5- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-two
Hydrogen quinazoline -3 (2H)-yl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) pentanamide (III-
2)；Its structural formula is as follows：
MS (ESI, m/z)：673.29[M-H]-
1H NMR (300MHz, DMSO) δ 11.00 (s, 1H), 9.75 (s, 1H), 7.80 (s, 1H), 7.49 (s, 2H), 7.34
(s, 3H), 7.21 (d, J=15.6Hz, 3H), 6.91 (d, J=7.2Hz, 1H), 6.80 (d, J=8.0Hz, 1H), 5.42 (s,
1H), 4.29 (d, J=34.4Hz, 2H), 3.52 (s, 1H), 3.29 (s, 2H), 2.99 (d, J=40.9Hz, 4H), 2.64 (s,
1H), 2.34 (s, 4H), 2.17 (s, 2H), 1.34 (s, 2H), 1.24 (s, 6H), 0.85 (s, 1H)
Embodiment 13：Prepare 6- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-two
Hydrogen quinazoline -3 (2H)-yl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) caproamide (III-
3)；Its structural formula is as follows：
MS (ESI, m/z)：687.30[M-H]-
1H NMR (300MHz, DMSO) δ 11.01 (s, 1H), 9.74 (t, J=17.6Hz, 1H), 7.87 (d, J=
44.1Hz, 1H), 7.51 (dd, J=27.1,21.2Hz, 3H), 7.23 (ddd, J=24.5,20.6,6.5Hz, 5H), 7.07-
6.65 (m, 2H), 5.41 (s, 1H), 4.48-4.22 (m, 2H), 3.86-3.40 (m, 3H), 3.24 (d, J=21.7Hz, 2H),
3.04-2.79 (m, 2H), 2.60 (d, J=17.9Hz, 1H), 2.34 (s, 3H), 2.17 (s, 2H), 1.94 (d, J=24.4Hz,
1H), 1.39-1.17 (m, 9H), 0.84 (d, J=6.1Hz, 2H)
Embodiment 14：Prepare 7- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-two
Hydrogen quinazoline -3 (2H)-yl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) heptamide (III-
4)；Its structural formula is as follows：
MS (ESI, m/z)：702.31[M-H]-
1H NMR (300MHz, DMSO) δ 11.01 (s, 1H), 9.74 (s, 1H), 7.80 (d, J=6.3Hz, 1H), 7.61-
7.41 (m, 2H), 7.40-7.18 (m, 7H), 7.03 (d, J=8.4Hz, 1H), 5.73 (s, 1H), 4.46-4.24 (m, 2H),
3.28 (s, 3H), 2.88 (dd, J=30.0,10.9Hz, 2H), 2.42-2.25 (m, 6H), 2.18 (s, 3H), 1.52 (d, J=
26.4Hz, 4H), 1.29 (dd, J=30.3,5.6Hz, 9H)
Embodiment 15：The protein level determination of activity of compound
The combination of first bromodomain domain of compound and BRD4 albumen n ends (hereinafter referred to as BRD4 (BD1)) is lived
Property test using AlphaScreen detection techniques.Compound primary dcreening operation concentration is 500nM.
HEPES buffer solution (50mM HEPES, 100mM NaCl, 0.1%BSA, 0.05%CHAPS, pH7.5) is prepared to be used for
Prepare BRD4 (BD1) albumen, Biotin mark histone H 4, testing compound (DMSO 0.1%), donor beads and
Acceptor beads solution.Take one piece of 384 orifice plate, according to arrangement, divide on plate testing compound hole, blank control wells (min,
Max), positive drug control wells.The μ L of compound solution 5 of various concentrations are separately added into testing compound hole and positive drug hole, it is empty
It is white to add the μ L (DMSO 0.1%) of buffer solution 5.Continue to add BRD4 (BD1) albumen to each hole in addition to blank control wells (min)
The μ L of solution 5, the μ L of buffer solution 5 are added to blank control wells (min).After being incubated 15 minutes at room temperature, Biotin marks are added per hole
The μ L of histone H 4 solution 5, continue at room temperature be incubated 1 hour after, add donor beads and acceptor beads solution
15 μ L, lucifuge are read glimmering with the Alpha mode (λ ex=680, λ em=570) of EnSpire detectors after being incubated at room temperature 1 hour
Light numerical value.
Numerical value processing：Inhibiting rate=(Max-Signal)/(Max-Min) × 100%
Wherein：Max：The value that the histone H 4 of Biotin marks is completely combined with albumen
Min：The histone H 4 background values of Biotin marks
Signal：Value under compound respective concentration
S curve is done with compound concentration and corresponding inhibiting rate.Obtain the IC of respective compound50。
The experimental result of part of compounds is shown in Table 1：
Inhibitory activity result of the preferred compound of table 1 to BRD4 (BD1).
| Compound | Inhibiting rate (%) 500nM | IC50(nM) |
| I-1 | 99.3 | 14 |
| I-2 | 100 | 15 |
| I-3 | 100 | 22 |
| I-4 | 100 | 22 |
| II-1 | 94.2 | 132 |
| II-2 | 89.3 | 148 |
| II-3 | 91.4 | 133 |
| II-4 | 98.3 | 99 |
| II-5 | 99.2 | 80 |
| II-6 | 100 | 68 |
| III-1 | 86.3 | 162 |
| III-2 | 90.7 | 143 |
| III-3 | 95.3 | 112 |
| III-4 | 99.6 | 72 |
Embodiment 16：Compound cellular level determination of activity
The Activity determination of Compound cellular level uses Celltiter-Glo fluorecyte viability examination methods.Will be in pair
Non-small cell lung cancer (NSCLC) A549, H1975 cell, colon cancer HT29, HCT15, HCT116 cell, the mammary gland in number growth period
Cancer MDA-MB-231, MDA-MB-468 cell and esophageal squamous cell carcinoma CaES-17 cells are seeded to 96 well culture plates, overnight incubation
Afterwards, 37 DEG C of testing compound, 5%CO are added272h is incubated, after terminating, balances measure reagent at room temperature within 30 minutes before detection.
30uL Celltiter-Glo reagents are added per hole, rock 96 orifice plate, 10 minutes inducing cell lysis.By 96 orifice plates at room temperature
2 minutes are incubated to stablize fluorescence signal.Fluorescence values are read using Envision detectors.
Numerical value processing：Inhibiting rate=(Max signal-Compound signal)/(Max signal-Min signal)
× 100%
Wherein：Max signal：The maximum read in DMSO effects
Min signal：The only minimum value of medium effect
Compound signal：Value under compound respective concentration
S curve is done with compound concentration and corresponding inhibiting rate.Obtain the IC of respective compound50。
The experimental result of part of compounds is shown in Table 2：
The preferred compound of table 2 to non-small cell lung cancer (NSCLC) A549, H1975 cell line, colon cancer HT29, HCT15,
The anti-increasing of HCT116 cell lines, breast cancer MDA-MB-231, MDA-MB-468 cell line and esophageal squamous cell carcinoma CaES-17 cell lines
Grow active (20uM, 72h).
Embodiment 17：Tablet
With containing compound in claim 1 (by taking the compound of embodiment 1 as an example) 10g, according to the general pressed disc method of pharmacy
After adding auxiliary material 20g to mix, 100 are pressed into, every weight 300mg.
Embodiment 18：Capsule
With containing compound in claim 1 (by taking the compound of embodiment 1 as an example) 10g, according to pharmacy capsule will
Ask after auxiliary material 20g is mixed, load Capsules, each capsule weight 300mg.
Embodiment 19：Injection
With containing compound in claim 1 (by taking the compound of embodiment 1 as an example) 10g, according to pharmacy conventional method, enter
Row charcoal absorption, after 0.65 μm of filtering with microporous membrane, insert nitrogen pot and hydro-acupuncture preparation is made, every fills 2mL, filling altogether
100 bottles.
Although describing the present invention by particular, modification and equivalent variations are for being proficient in this field
It will be apparent from for technical staff, and they are included within the scope of the invention.
Claims (7)
1. difunctional small molecule or its pharmaceutically acceptable salt, hydrate or prodrug shown in below formula：
A-L-B
Wherein：
A is the smaller ligand of cereblon albumen in E3 SCF-complexes, includes amides compound, phthalyl
Imine compound, Thalidomide or derivatives thereof, lenalidomide or derivatives thereof, pomalidomide or derivatives thereof, partly matches somebody with somebody
Body general structure is as follows：
Wherein：
W is selected from CH2, C=O, SO2, NH, N- alkyl；
X is selected from O, S；
Y is selected from NH, N- alkyl, N- aryl, N- heterocycles, N- cycloalkyl, O, S；
Z be selected from-alkyl ,-cycloalkyl ,-Cl ,-F；
G, G ' is selected from-H ,-alkyl ,-OH ,-CH2- heterocycle；
Q1、Q2、Q3、Q4Selected from C, N, NO.
L is linking arm, comprising non-linear chain, aliphatic chain, fragrant chain, Heteroaryl ring structures chain, is connected by covalent bond with A and B,
Shown in part connection arm configuration below formula：
Wherein：
N is selected from the integer between 1-10.
B is BET protein inhibitors or derivatives thereof.
2. according to the compound below formula (Ia), (IIa) and (IIIa) described in claim 1,
Wherein：
R1Selected from-H ,-D ,-CH3、-CH2F、CHF2、-CF3、-CH2D、-CHD2、-CD3、-CH2CH3、-CH2CH2CH3；
R2、R3It is identical or different, separately selected from-H ,-D ,-F ,-Cl ,-Br ,-I ,-NO2、-CN、-NH2、-OH、-CH3、-
CH2F、CHF2、-CF3、-CH2D、-CHD2、-CD3、-CH2CH3；
N is selected from the integer between 1-10.
3. according to the compound described in claim 2, wherein：
R1Selected from-H ,-CH3、-CH2CH3；
R2、R3It is identical or different, separately selected from-H ,-F ,-Cl ,-Br ,-CH3、-CH2F、CHF2、-CF3、-CH2D、-
CHD2、-CD3；
N is selected from the integer between 1-6.
4. according to the compound described in claim 1-3, wherein, the compound is one of following compounds or it is acceptable
Salt, hydrate or prodrug,
Ia class compounds：
2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxos
Isoindoline -4- bases) acetamide；
2- (2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydro quinoline azoles
Quinoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl) ethyoxyl)-N- (2- (2,6- dioxopiperidine -3-
Base) -1- oxoisoindolines -4- bases) acetamide；
2- (2- (2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-dihydro quinoline azoles
Quinoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl) ethyoxyl) ethyoxyl)-N- (2- (2,6- dioxos
Piperidines -3- bases) -1- oxoisoindolines -4- bases) acetamide；
2- (2- (2- (2- (2- (4- ((6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-dihydro quinoline
Oxazoline -3 (2H)-yl) methyl) -1H-1,2,3- triazole -1- bases) ethyoxyl) ethyoxyl) ethyoxyl) ethyoxyl)-N- (2-
(2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) acetamide；
IIa class compounds：
3- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) propionamide；
4- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) butyramide；
5- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) pentanamide；
6- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) caproamide；
7- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) heptamide；
8- (2- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazoline -3
(2H)-yl) acetylamino)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) caprylamide；
IIIa class compounds
3- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazolines -3 (2H) -
Base)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) propionamide；
5- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazolines -3 (2H) -
Base)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) pentanamide；
6- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazolines -3 (2H) -
Base)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) caproamide；
7- (6- (3,5- dimethyl isoxazole -4- bases) -1- methyl -2- oxos -4- phenyl-Isosorbide-5-Nitrae-dihydroquinazolines -3 (2H) -
Base)-N- (2- (2,6- dioxopiperidine -3- bases) -1- oxoisoindolines -4- bases) heptamide.
5. the preparation method of the compound any one of claim 1-4, is prepared by following methods：
The compounds process for production thereof of formula (Ia), method are as follows：
Step a：Compound I reacts to obtain compound II with paratoluensulfonyl chloride；
Step b：Compound II reacts to obtain compound III with Sodium azide；
Step c：Compound III reacts to obtain compound IV with bromo-acetic acid tert-butyl；
Step d：Compound IV reacts to obtain compound V with trifluoroacetic acid；
Step e：Compound V reacts to obtain compound VI with thionyl chloride；
Step f：Compound VI and lenalidomide or derivatives thereof reaction obtain compound VII；
Step g：Compound VIII and VII reacts to obtain the compound of formula (Ia) expression；
The compounds process for production thereof of formula (IIa), method are as follows：
Step h：Compound IX and lenalidomide or derivatives thereof reaction obtain compound X；
Step i：Compound X reacts to obtain compound XI with trifluoroacetic acid；
Step j：Compound XII and XI reacts to obtain the compound of formula (IIa) expression；
The compounds process for production thereof of formula (IIIa), method are as follows：
Step k：Compound XIII and XVI reacts to obtain compound XV；
Step l：Compound XV and lenalidomide or derivatives thereof reaction obtain the compound of formula (IIIa) expression；
Wherein, R1、R2、R3It is as defined above with n.
6. a kind of pharmaceutical composition include claim 1-4 any one described in compound and its pharmaceutically acceptable salt or
Pharmaceutically acceptable carrier, excipient, diluent, assistant agent, medium, or combinations thereof.
7. the compound of any one and its pharmaceutically acceptable salt or the combination described in claim 6 in claim 1-4
Application of the thing in the medicine for treating or preventing cancer is prepared.
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